title: The primary DNA-binding subsite of the rat pol β. Energetics of interactions of the 8-kDa domain of the enzyme with the ssDNA.
abstract: Interactions of the 8-kDa domain of the rat pol β and the intact enzyme with the ssDNA have been studied, using the quantitative fluorescence titration technique. The 8-kDa domain induces large topological changes in the bound DNA structure and engages much larger fragments of the DNA than when embedded in the intact enzyme. The DNA affinity of the domain is predominantly driven by entropy changes, dominated by the water release from the protein. The thermodynamic characteristics dramatically change when the domain is embedded in the intact polymerase, indicating the presence of significant communication between the 8-kDa domain and the catalytic 31-kDa domain. The diminished water release from the 31-kDa domain strongly contributes to its dramatically lower DNA affinity, as compared to the 8-kDa domain. Unlike the 8-kDa domain, the DNA binding of the intact pol β is driven by entropy changes, originating from the structural changes of the formed complexes.
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Texas Medical Branch at Galveston, 77555-1053, United States.
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Jezewska
authors.forename: Michal R
authors.initials: MR
authors.lastname: Szymanski
authors.forename: Wlodzimierz
authors.initials: W
authors.lastname: Bujalowski
mesh_terms: D000818:Animals; D001665:Binding Sites; D019951:DNA Polymerase beta / Q000737:chemistry* / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000737:chemistry* / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry; D008274:Magnesium / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D051381:Rats; D012997:Solvents / Q000737:chemistry; D013696:Temperature; D013816:Thermodynamics
chemical_list: D004277:DNA, Single-Stranded; D005456:Fluorescent Dyes; D012997:Solvents; D019951:DNA Polymerase beta; D008274:Magnesium